Confocal Microscopy and Image Processing Techniques for Online Monitoring of Engineered Tissue

Evaluation of engineered tissue is often limited to endpoint analyses, such as characterizing histology, gene expression and solutes [1]. Most of these applied analysis approaches are based on immunochemistry procedures that require excision, fixation and sectioning of the tissue as well as cell membrane perforation and labeling of proteins [2]. These analyses are time-consuming, do not facilitate high throughput processing and do not allow for online monitoring of engineered tissue. Because of these limitations, there is a need for online, high throughput monitoring techniques to evaluate engineered tissue. In this work, we introduce an approach for microscopic imaging and online analysis of living engineered tissue. The approach is based on application of a non-toxic dye specific for the extracellular space and subsequent interrogation by in vivo confocal microcopy. We hypothesized that the approach will allow for online characterization of cell structure.